製品の概要

This antibody detects calmodulin. It does not detect parvalbumin, tropinin, S-100, or myosin light chain kinase (MLCK).By Western blot, this antibody detects a 17 kDa protein representing calmodulin from Dictyostelium cell lysate. Immunohistochemical staining of calmodulin in Dictyostelium cells with this antibody results in staining of the contractile vacuoles.

製品の特性

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

バッファー

Preservative: 0.05% Sodium azideConstituent: 99% PBS

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精製度

Protein A purified

一次抗体 備考

Calmodulin is a small, highly conserved calcium binding protein found in all eukaryotic cells. With the capacity to bind up to four calcium ions, this 17 kDa protein acts as an important intracellular receptor for regulatory calcium signals. As it binds calcium, calmodulin undergoes conformational changes which can increase its affinity for target proteins. It acts both directly, through interaction with key target enzymes, and indirectly, via specific kinases.
Studies have found that calmodulin participates in the regulation of several biological processes including energy and biosynthetic metabolism, cell motility, exocytosis, cytoskeletal assembly, and intracellular modulation of both cAMP and calcium concentrations.

アプリケーション

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション

Abreviews

特記事項

Flow Cyt

Use 2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA

Use at an assay dependent concentration.

WB

1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).

IHC-P

1/20.

ICC

1/50.

IP

Use at an assay dependent concentration.

ICC/IF

1/20.

ターゲット情報

関連性

Function: Calmodulin mediates the control of a large number of enzymes and other proteins by Ca(2+). Among the enzymes to be stimulated by the calmodulin-Ca(2+) complex are a number of protein kinases and phosphatases. Together with CEP110 and centrin, is involved in a genetic pathway that regulates the centrosome cycle and progression through cytokinesis.

There are three genes which encode an identical calcium binding protein which is one of the four subunits of phosphorylase kinase.

別名

CALM 1 antibody

CALM 2 antibody

CALM 3 antibody

CALM antibody

CALM1 antibody

CALM2 antibody

CALM3 antibody

CALML2 antibody

calmodulin 1 (phosphorylase kinase, delta) antibody

Calmodulin 1 antibody

Calmodulin 2 (phosphorylase kinase, delta) antibody

Calmodulin 2 antibody

Calmodulin 3 (phosphorylase kinase, delta) antibody

Calmodulin 3 antibody

CAM 2 antibody

CAM 3 antibody

CAM I antibody

CAM1 antibody

CAM2 antibody

CAM3 antibody

CAMB antibody

CAMC antibody

CAMI antibody

CAMII antibody

CAMIII antibody

CPVT4 antibody

DD132 antibody

FLJ99410 antibody

LP7057 protein antibody

PHKD antibody

PHKD2 antibody

PHKD3 antibody

phosphorylase kinase delta antibody

phosphorylase kinase, delta subunit antibody

see all

画像

Flow Cytometry - Anti-Calmodulin antibody [2D1] (ab2860)

Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of PC12 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Flow Cytometry - Anti-Calmodulin antibody [2D1] (ab2860)

Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of MCF-7 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at a dilution of 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Flow Cytometry - Anti-Calmodulin antibody [2D1] (ab2860)

Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of C6 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in HeLa cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in A2058 melanoma cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in C6 glioma cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Immunohistochemistry was performed on normal biopsies of deparaffinized Rat testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on normal biopsies of deparaffinized Rat cerebellum tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Thank you for your phone enquiry and questionaire.
I'm sorry to hear you are having a problem with ab2860 (calmodulin Ab) in Western blotting. We have received other enquiries regarding this antibody and I would like to suggest the following modifi...

Thank you for your enquiry.
I browsed the reference, Cell Mot. Cytoskel., 18: 113-122, 1991. There are a few suggestions, taken from the article, that I can give. These researchers had difficulty with their proteins transferring through the memb...

I'm very sorry to hear you are experiencing problems with ab2860.
I would also expect the lysates in rat and E.coli samples to work with this antibody, however there may be several reasons for the lack of signal you are currently experiencing.
...

Thank you for those extra details about your protocol this helped me a lot as I looked deeper into the cross reactivity of this antibody with your samples: 293T and HeLa cells are from human origin and 3T3 cells are from mouse origin.
This could be ...

I'm sorry to hear you are having a problem with ab2860. We have not had a complaint about this antibody before.
Thank you for taking the time to fill in our questionnaire, it helped me to understand your problem and it is useful to know that another...

Thank you for your enquiry and for your patience. Regarding the specific epitope mapping, we have not performed these studies. Both immunogens were: Calmodulin purified from Dictyostelium discoideum and both antibodies have been tested for application ...